RESUMO
Encephalomyocarditis virus (EMCV) infections can cause losses in pig farms all over the world. Rapid, sensitive and unequivocal detection of this virus is therefore essential for the diagnosis and control of the disease. An RT-PCR assay was developed, optimized and evaluated for encephalomyocarditis virus detection in organ based on a pair of primers that amplifies a 165 bp DNA fragment from a highly conserved nucleotide region of the viral 3D glycoprotein. PCR products of the expected size were obtained from Cuban EMCV 744/03 strain. Non-specific reactions were not observed when other porcine RNA genome viruses and uninfected cells were used. The analytical sensitivity of the test was estimated to be 2 TCID50/50 mL. The analysis of tissue homogenate samples from naturally infected animals proved the potential usefulness of the method for a rapid disease diagnosis from field cases.
Assuntos
Animais , Genoma/genética , Técnicas In Vitro , Nucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vírus de RNA , Suínos , Vírus da Encefalomiocardite/genética , Vírus da Encefalomiocardite/isolamento & purificação , Métodos , Técnicas de Amplificação de Ácido Nucleico , MétodosRESUMO
GDP-mannose-3', 5' -epimerase (GME), which converts GDP-mannose into GDP-L-galactose, is essential for the biosynthesis of L-ascorbic acid in higher plants. The molecular characterization of two GME genes from rice has been reported. Firstly,both cDNAs were isolated from the rice mature leaves using RT-PCR technique. By comparing their sequences with homologues from other plants, it was found that GME genes were highly conserved among plant species, though phylogenetic study showed that all known GMEs could be divided into two distinct groups corresponding to monocots and dicots. Secondly, the genomic organization of rice OsGME genes was investigated, and a similarity of splice patterns was revealed. Finally, the expression patterns of the two cDNAs have been studied in various tissues and under different stress conditions by semiquantitative RT-PCR assay. The results showed that the OsGME1 transcript was up-regulated in response to cold stress, and gibberellin might regulate L-ascorbic acid levels by affecting transcription of both OsGME genes.